Bivalent virus vaccine, containing rubella TCRB-19 strain and mumps NK-M46 strain (MR vaccine), was administered to a total of 95 healthy children who had already received measles vaccine or had been infected with wild measles virus. The seroconversion rates for rubella and mumps viruses in subjects having no antibody to rubella or to mumps virus were 99%(75/76) and 97%(63/65), respectively, at 6-8 weeks after vaccination. The seroconversion rates for both rubella and mumps in vaccinees initially seronegative to both viruses were 95%(56/59). Immune responses after MR vaccine injection were comparable to those after administration of monovalent rubella or mumps vaccine. Clinical reactions observed in some subjects who received MR vaccine were mild fever (3.6%), examthem (8%), lymphadenopathy (1.8%), and swelling of the parotis region (1.8%). MR vaccine could be simultaneously injected with varicella vaccine at the opposite site producing no adverse effect on immune response. Our results indicate that MR vaccine is a safe and effective vaccine, especially for children who have had wild measles or who have received measles vaccine.

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The possibility of rapid diagnosis of Mycoplasma pneumoniae infection by immunobinding assay is described. Immunobinding assay which was developed by Kotani and McGarrity is a simple and rapid method for identification of mycoplasmas. Small amounts of antigen were spotted onto the nitrocellurose membrane. It was treated with a specific rabbit antisera against M. pneumoniae. The antigen-antibody complex was visualised with the avidin-biotin horseradish peroxidase method. Cross reaction was seen between M. pneumoniae and M. genitalium. Throat swabs from hamsters infected with M. pneumoniae were positive on 7th and 14th day after infection.
Although the cross reaction was seen between M. pneumoniae and M. genitalium, this method could be useful for rapid diagnosis of M. pneumoniae infection.

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It is well known that the etiologic agent, Coxiella burnetii, exhibits an antigenic phase variation (phase I to phase II), and the diagnostic significance of the relative antibody titers against phase I and phase II antigens is pointed out. Therefore both phase I and phase II antigens are necessary for the serological examination of Q fever. But it is not so easy to prepare and maintain the phase II antigen by the conventional method. In the present study we tried to prepare the phase II antigen for immunofluorescence test by chemical treatment of the phase I antigen. As a result, treatment of the TK-1 strain of C. burnetii (phase I) with 10% trichloroacetic acid for 4 hr at 4°C modified the antigenicity. The modified antigen reacted strongly with anti-phase II antibody.

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Rhinoviruses (HRVs) were isolated from 307 children (7.1%) in the virological serveillance of 4334 children with acute respiratory tract illnesses in Morioka, Japan (September 1973-December 1983).
Although HRVs were isolated throughout the year, frequency of HRV infection was significantly higher (p<0.001) during the April-November (233/2853; 8.2%) than during the December-March (47/1481; 5.0%). There were two peaks of incidence in May (9.5%) and September (9.1%). During the May-September, the rate of HRV infection was higher in patients under the age of 11 months than the next higher group of 1-2 years old (p<0.001). The incidence decreased with increasing age.
The illnesses of HRV infection were analysed in 294 patients, except one patient who had symptoms of measles, from whom HRV was isolated singly. Although HRV-associated illnesses were generally mild (57.5%). Upper respiratory tract illnesses (URTIs) with fever were found in 22.1% and lower respiratory tract illnesses (LRTIs) in 20.4% of these. The rate of LRTI was higher during the epidemic period (April-September) than other periods (p<0.02).
Major symptoms of HRV-associated illnesses observed were sore throat (87.4%), cough (84.0%), and nasal obstruction and/or discharge (72.8%). Wheezing was observed in 21.8% of these.
From 19 (21.8%) of 47 patients clinically diagnosed as asthmatic bronchitis in this servey, viruses were isolated. HRV was detected most frequently in 12.8% of these patients, followed by respiratory syncytial virus (RSV, 6.4%) and adenovirus (2.1%). HRV-and RSV-associated asthmatic bronchitis were observed during April-September and November-February, respectively.
Viral dual infections were detected in total 20 cases included 12 HRV-associated cases. In no case was the illness of greater severity than might have been caused by either agent acting singly.

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During a 20-year period disseminated mycobacteriosis occurred in 11 (1.1%) of a total of 1006 patients with severe hematologic disorders, with the frequency remaining almost unchanged. The diagnosis in three patients (27%) was made only at autopsy. Tuberculosis accounted for 64% of all cases. Female preponderance was seen with a male-to-female ratio of 3: 8. The major factors associated with dissemination included immunosuppression, weight loss, old age, and diabetes mellitus. Fever was the most common clinical symptom. Chest X-ray abnormalities, hypoproteinemia, liver dysfunction, and hypoxemia were noted in most cases. The prognosis of tuberculosis depended mainly on early diagnosis and treatment, while that for the nontuberculous variety was largely influenced by the underlying disease. Thus, our findings indicated that clinicians must suspect disseminated mycobacteriosis especially in any febrile patient with recent pulmonary pathology on chest X-ray, so that an adequate trial of therapy can be provided.

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Induction to cell wall-deficient bacteria has been suggested to be a cause of intractable and opportunistic infection after chemotherapy. Spheroplast formation by β-lactam antibiotic in not so high osmotic pressure was investigated in six species of gram-negative bacteria. Some species of gram-negative bacteria were induced to form spheroplast at a high rate by 1: 10 concentration of ceftizoxime in the presence of Ca²⁺ or Mg²⁺. Especially in 38% of Prosteus mirabi is and P. vulgaris, more than 40% of the original cells were induced to form spheroplast by ceftizoxime in a medium supplemented with 40mM Ca²⁺. The same formation rate was also found in 22% of Serratia marcescens. Formation rates in the drug sensitive strains of S. marcescens were enhanced as the drug concentration increased. Ca²⁺ was more effective in spheroplast formation than Mg²⁺.

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In conjunction with the Tokyo Branch of Japan Association for Maternal Welfare, we have been carrying out examinations of Chlamydial infection on women, especially pregnant women, living in Tokyo.
Specimens were collected from 212 gyneco-obstetric clinics in Tokyo during 4 years from January 1987-December 1990. A total of 13, 925 swab specimens from patients who were suspected of sexually transmitted diseases from clinical findings and pregnant women who requested the examination were tested for the presence of antigen to C. trachomatis with EIA (Chlamydiazyme ^®).
Epidemiological analysis based on the results and case cards that were described sex, age, occupation and clinical findings were conducted. The results obtained are briefly summarized as follows.
1) C. trachomatis antigen was detected in 12.8%(1, 237/9, 657) of the female patients, 31.0%(124/400) of the male patients and 6.8%(10/168) of the infants. The detection rates of C. trachomatis antigen in the male patients was significantly higher than that in the female and the infants patients. C. trachomatis antigen was detected in 6.1%(226/3, 683) of pregnant women.
2) The detection rates of C. trachomatis antigen were compared by occupation of female patients. Most high rate was 26.6%(53/199) in bar hostesses, subsequently 20.1%(102/508) in students, 19.8%(59/298) in prostitutes, 13.6%(455/3, 348) in office girls and 7.2%(471/6, 573) inhouse-wives. It was noteworthy that the detection rates of C. trachomatis antigen in students was as similally high as hat in prostitutes.
3) The detection rates of C. trachomatis antigen was also compared by age groups. The detection rate had a tendency to be high in younger people. The rates in late teens to early 20s were 19.9%(642/3, 221) in the female patients, 11.3%(85/750) in the pregnant women, respectively.
On the other hand, the rates in the late 20s and abobe were 8.9%(549/6, 140) in the former, 4.4%(136/2, 847) in the latter, respectively. In both groups, there was a significant difference between the detection rates of C. trachomatis antigen in subjects in their late teens to early 20's and that in subjects in their late 20s and above (p<0.01).

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DNA probe assay kit for the identification of Mycobacterium tuberculosis was evaluated. This kit isbased on beads capture method and using a 20 well assay tray.
The culture isolate is suspended with 0.5 ml sterilized water in the tube containing φ3mm glassbeads for dispersion, transferred to a well of the assay tray. After lysation and adsorption of the nucleic acids to the capture bead, ¹²⁵I-DNA probe specific for the M. tuberculosis is added to the sample and hybridized for 1 hour at 65°C. Hybridized probe trapped on the capture bead is quantitated using agamma counter.
This hybridization assay kit can detect more than 1×10⁵ bacteria per assay. Using the contr DNA (synthesized oligonucleotide complementary to the probe sequences) and suspension of the culture isolates, the intra assay C. V. value was 4.3% and 5.2% respectively.
To compare this probe assay kit with the conventional culture identification method, a total of 144 culture isolates were examined. This test for the M. tuberculosis had 99.3% agreement with the conventional identification procedures, and demonstrated 100% sensitivity and 98.2% specificity.
The suspension of culture isolates can be stocked by freezing, the samples can be assayed together when they have accumulated instead of every day. As shown in the above, this assay kit demonstrates a high level of specificity and sensitivity. This test is very easy to perform, because it is not necessar to prepare any instrument for the extraction of the nucleic acids from the bacteria and there is nothing to complicate the procedure, in addition, results are available in less than 3 hours.
In this case, because the assay kit employs the bead captured solid phase methods, there seems to be many advantages in using the automatic assay system.

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We determined the minimum inhibitory concentrations (MICs) of rokitamycin (TMS-19-Q, RKM), a macrolide antimicrobial agent, against strains of various bacterial species isolated from enteritis patients, and compared them with those of josamycin (JM), erythromycin (EM) and ofloxacin (OFLX). MIC₉₀ of RKM against 147 strains of Campylobacter jejuni, and each 25 strains of Shigella spp., Salmonella spp. and diarrheagenic Escherichia coli were 1.56, 200, 800 and 200μg/ml, respectively. There was only one RKM resistant (MIC>100μg/ml)C. jejuni strain, while most of the strains of the other species were resistant to RKM. MIC values of the other drugs were all similar to those of RKM. MIC₉₀ of OFLX against 147 strains of C. jejuni was 0.78μg/ml, lower than other drugs.

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We compared the results obtained with the polymerase chain reaction (PCR) and virus isolation from peripheral blood mononuclear cells (PBMC) in HIV seropositive and seronegative persons. Three primer pairs of SK38/39 (gag). SK29/30 (LTR) and SK68/69 (env) were used in the amplification of the HIV DNA sequences, and KM29/38 (β-globin) was used as the inner control. The PCR-positive rate among the virus-isolation-positive persons was SK38/39: 100%(22/22), SK29/30: 95.5%(21/22) and SK68/69: 90.0%(20/22). The PCR-positive rate among the virus-isolation-negative persons was SK38/39: 60%(6/10), SK29/30: 60%(6/10) and SK68/69: 80%(8/10), and two subjects were PCRnegative with all primer pairs. We could not detect HIV DNA from seronegative samples, and all subjects were positive with the inner control.
Each primer pair expressed a different PCR-positive rate. There are possible explanations for the low PCR-negative rate on virus-isolation negative-subjects that the number of infected cell was rare or infected HIV contained genetic variations or deletions. We considered that the results of PCR correlated with the character of HIV as infectivity.

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It is well known that long-term administration of erythromycin (EM) at a small dose is effective for persistent infections with Pseudomonas aeruginosa in diffuse panbronchiolitis or chronic bronchitis patients. Since EM is less active against P. aeruginosa in vitro, we have been interested in the mechanisms of clinical efficacy of EM in these patients. This study examines the effect of macrolide antibiotics on human serum-bactericidal sensitivity of P. aeruginosa S-6, clinically isolated from the patient with respiratory tract infection.
A significant increase in serum-bactericidal sensitivity of P. aeruginosa S-6 was observed on agar containing EM of 10μg/ml after incubation for 36-60 hours (p<0.05). The enhancement of serum sensitivity of P. aeruginosa S-6 was apparently observed even at a concentration of EM 1.5μg/ml after the 48 hours incubation (p<0.01). Of other macrolide antibiotics used, clarithromycin (CAM) also increased the serum-bactericidal sensitivity of P. aeruginosa S-6 as well as EM, however no change in the sensitivity was found with kitasamycin, josamycin, rokitamycin and oleandomycin.
The reuslts suggest that the change of serum-bactericidal sensitivity of P. aeruginosa induced by EM or CAM may, in part, contribute to the clinical efficacy of these antibiotics against persistent pulmonary P. aeruginosa infections.

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Etiology of bacterial infections in the field of oral surgery was studied. A total of 270 samples collected from patients with encapsulated abscess in their oral cavities was examined and bacteria were isolated from the 244 samples (90.4%). The following results were found;
1) Organisms more than one from one sample were frequently isolated from cases with parodontitis, pericoronitis and gnathitis. Isolation of anaerobic bacteria was common (54.2%).
2)Streptococcus milleri and Streptococcus sanguis and Capnocytophaga species were the most common isolates among aerobic gram-positive and gram-negative bacteria, respectively.
3) Peptostreptococcus micros and Eubacterium lentum were most frequent isolates among gramnegative anaerobic bacteria. Among gram-negative bacteria, Oral Group Bacteroides, especially Bacteroides gingivalis, Bacteroides intermedius, Bacteroides buccae and Bacteroides oralis were most prominent.
4) Isolation frequency of bacteria (both species and strains) was high from samples obtained from patients before antibiotic chemotherapy.
5) Most strains were sensitive to Midecamycin acetate and Josamycin. Minimum inhibitory concentration of 80% isolates (MIC₈₀) against these antibiotics was 0.39μg/ml.

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Ureaplasma urealyticum was investigated in urine from 765 outpatients who visited Jikei University Affiliated Hospital and Tokyo Metropolitan Taito Hospital from June, 1988 to December, 1989 in order to clarify the pathogenicity of U. urealyticum in human genitourinary tract.
U. urealyticum in urine was detected by means of Taylor-Robinson's method.
The positive rates of U. urealyticum were 31.5% in 146 patients with gonococcal urethritis, 33.8% in 334 patients with non-gonococcal urethritis, 17.5% in non-bacterial chronic prostatitis and 27.5% in the other patients without infectious diseases, respectively; no significant difference was seen among these groups. U. urealyticum was detected in the urine from 32.1% of the 28% patients who were younger than 12. However, U. urealyticum was detected in the urine from 5.6% of the 18 patients who were older than 70. Therefore, there was no relationship between the age and U. urealyticum-positive rate in urine. Furthermore, there was no relationship between the detection of U. urealyticum and the subjective and objective findings in the patients with urethritis before and after the treatment
From these results, it is presumed that U. urealyticum has no pathogenicity in human genitourinary tract.

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We reported a case of salmonellosis treated with ofloxacin (OFLX) which showed excellent clinical and bacteriological effect in a 22 year-old Japanese male with Salmonellosis paratyphi A.
He had stayed in India from Sept. 6, 1990 to Oct. 13, 1990. On Oct. 25, 1990, he complained of a high fever and headache. On Oct. 29, he was admitted to our hospital and was diagnosed as Salmonellosis paratyphi A by the blood culture. He was treated with 2.0g/day of chloramphenicol (CP) for 7 days, but the clinical efficacy was not sufficent. Therefore, we added 900mg/day of OFLX for 10 days. He was treated successfully with them, the temperature became on the 2nd day.
No side effect and no changes of laboratory data were observed and no recurrence was observed clinically and bacteriologically for three months after his discharge.

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We report a case of sepsis who died caused by Pasteurella multocida subsp.multocide sepsis. A 68-year-old male was admitted to Azusawa Hospital because of disturbance of consciousness. He had been suffering from diabetes mellitus combined with gangrene, but received no treatment. The patient died 24h after hospitalization, and Pasteurella multocida subsp.multocida was isolated from hi blood. Laboratory tests showed that CRP; 5+WBC; 15400/μl, TP; 5.2g/dl. Although Pasteurella multocida subsp. multocida seemed to cause mild infection in healthy subjects, it can cause sever systemic illnesses such as sepsis and meningitis in compromised hosts. It should be considered that the contact with pets will increase the incidence of systemic sever infection with this agents.

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We encountered a 14-year-old male patient with a destructive abscess of nasal septum, caused by Streptococcus milleri. He was successfully treated with Clindamycin in combination with surgical intervention.
We emphasized the significance of Streptococcus milleri as a causative agent for abscess formation, and clindamycin should be considered as a first choice of antibiotics against Streptococcus milleri infection.

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